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Sputum Processing Method for Lateral Flow Immunochromatographic Assays to Detect Coronaviruses

  • Aram Kang (College of Pharmacy, Korea University) ;
  • Minjoo Yeom (College of Pharmacy, Korea University) ;
  • Hyekwon Kim (Department of Microbiology, Chungbuk National University) ;
  • Sun-Woo Yoon (Korea Research Institute of Bioscience and Biotechnology) ;
  • Dae-Gwin Jeong (Korea Research Institute of Bioscience and Biotechnology) ;
  • Hyong-Joon Moon (College of Healthcare & Biotechnology, Semyung University) ;
  • Kwang-Soo Lyoo (Korea Zoonosis Research Institute, Chonbuk National University) ;
  • Woonsung Na (College of Veterinary Medicine, Chonnam National University) ;
  • Daesub Song (College of Pharmacy, Korea University)
  • Received : 2020.10.04
  • Accepted : 2021.01.30
  • Published : 2021.02.28
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Abstract

Coronavirus causes an infectious disease in various species and crosses the species barriers leading to the outbreak of zoonotic diseases. Due to the respiratory diseases are mainly caused in humans and viruses are replicated and excreted through the respiratory tract, the nasal fluid and sputum are mainly used for diagnosis. Early diagnosis of coronavirus plays an important role in preventing its spread and is essential for quarantine policies. For rapid decision and prompt triage of infected host, the immunochromatographic assay (ICA) has been widely used for point of care testing. However, when the ICA is applied to an expectorated sputum in which antigens are present, the viscosity of sputum interferes with the migration of the antigens on the test strip. To overcome this limitation, it is necessary to use a mucolytic agent without affecting the antigens. In this study, we combined known mucolytic agents to lower the viscosity of sputum and applied that to alpha and beta coronavirus, porcine epidemic diarrhea virus (PEDV) and Middle East respiratory syndrome coronavirus (MERS-CoV), respectively, spiked in sputum to find optimal pretreatment conditions. The pretreatment method using tris(2-carboxyethyl)phosphine (TCEP) and BSA was suitable for ICA diagnosis of sputum samples spiked with PEDV and MERS-CoV. This sensitive assay for the detection of coronavirus in sputum provides an useful information for the diagnosis of pathogen in low respiratory tract.

Keywords

Acknowledgement

This work was supported by a Korea University grant.

References

  1. Yamamoto M, Matsuyama S, Li X, Takeda M, Kawaguchi Y, Inoue JI, Matsuda Z. Identification of nafamostat as a potent inhibitor of middle east respiratory syndrome coronavirus S protein-mediated membrane fusion using the split-protein-based cell-cell fusion assay. Antimicrob Agents Chemother 2016;60:6532-6539. https://doi.org/10.1128/AAC.01043-16
  2. van Boheemen S, de Graaf M, Lauber C, Bestebroer TM, Raj VS, Zaki AM, Osterhaus AD, Haagmans BL, Gorbalenya AE, Snijder EJ, et al. Genomic characterization of a newly discovered coronavirus associated with acute respiratory distress syndrome in humans. MBio 2012;3:e00473-12.
  3. Annan A, Baldwin HJ, Corman VM, Klose SM, Owusu M, Nkrumah EE, Badu EK, Anti P, Agbenyega O, Meyer B, et al. Human betacoronavirus 2c EMC/2012-related viruses in bats, Ghana and Europe. Emerg Infect Dis 2013;19:456-459. https://doi.org/10.3201/eid1903.121503
  4. Omrani AS, Al-Tawfiq JA, Memish ZA. Middle East respiratory syndrome coronavirus (MERS-CoV): animal to human interaction. Pathog Glob Health 2015;109:354-362. https://doi.org/10.1080/20477724.2015.1122852
  5. World Health Organization. Middle East respiratory syndrome coronavirus (MERS-CoV) [Internet]. Available at https://www.who.int/emergencies/mers-cov/en/ [accessed on 28 September 2020]. 
  6. Korea Centers for Disease Control and Prevention. Middle East respiratory syndrome coronavirus outbreak in the Republic of Korea, 2015. Osong Public Health Res Perspect 2015;6:269-278. https://doi.org/10.1016/j.phrp.2015.08.006
  7. Huang P, Wang H, Cao Z, Jin H, Chi H, Zhao J, Yu B, Yan F, Hu X, Wu F, et al. A rapid and specific assay for the detection of MERS-CoV. Front Microbiol 2018;9:1101.
  8. Shirato K, Semba S, El-Kafrawy SA, Hassan AM, Tolah AM, Takayama I, Kageyama T, Notomi T, Kamitani W, Matsuyama S, et al. Development of fluorescent reverse transcription loop-mediated isothermal amplification (RT-LAMP) using quenching probes for the detection of the Middle East respiratory syndrome coronavirus. J Virol Methods 2018;258:41-48. https://doi.org/10.1016/j.jviromet.2018.05.006
  9. Shirato K, Yano T, Senba S, Akachi S, Kobayashi T, Nishinaka T, Notomi T, Matsuyama S. Detection of Middle East respiratory syndrome coronavirus using reverse transcription loop-mediated isothermal amplification (RT-LAMP). Virol J 2014;11:139.
  10. Bhadra S, Jiang YS, Kumar MR, Johnson RF, Hensley LE, Ellington AD. Real-time sequence-validated loop-mediated isothermal amplification assays for detection of Middle East respiratory syndrome coronavirus (MERS-CoV). PLoS One 2015;10:e0123126.
  11. Al Johani S, Hajeer AH. MERS-CoV diagnosis: an update. J Infect Public Health 2016;9:216-219. https://doi.org/10.1016/j.jiph.2016.04.005
  12. Chen Y, Chan KH, Kang Y, Chen H, Luk HK, Poon RW, Chan JF, Yuen KY, Xia N, Lau SK, et al. A sensitive and specific antigen detection assay for Middle East respiratory syndrome coronavirus. Emerg Microbes Infect 2015;4:e26.
  13. Song D, Ha G, Serhan W, Eltahir Y, Yusof M, Hashem F, Elsayed E, Marzoug B, Abdelazim A, Al Muhairi S. Development and validation of a rapid immunochromatographic assay for detection of Middle East respiratory syndrome coronavirus antigen in dromedary camels. J Clin Microbiol 2015;53:1178-1182. https://doi.org/10.1128/JCM.03096-14
  14. Takano T, Hohdatsu T. Serological diagnosis of feline coronavirus infection by immunochromatographic test. Methods Mol Biol 2015;1282:33-39. https://doi.org/10.1007/978-1-4939-2438-7_3
  15. Al Hammadi ZM, Chu DK, Eltahir YM, Al Hosani F, Al Mulla M, Tarnini W, Hall AJ, Perera RA, Abdelkhalek MM, Peiris JS, et al. Asymptomatic MERS-CoV infection in humans possibly linked to infected dromedaries imported from Oman to United Arab Emirates, May 2015. Emerg Infect Dis 2015;21:2197-2200. https://doi.org/10.3201/eid2112.151132
  16. Lee JH, Lee CS, Lee HB. An Appropriate lower respiratory tract specimen is essential for diagnosis of Middle East respiratory syndrome (MERS). J Korean Med Sci 2015;30:1207-1208. https://doi.org/10.3346/jkms.2015.30.8.1207
  17. Guery B, Poissy J, el Mansouf L, Sejourne C, Ettahar N, Lemaire X, Vuotto F, Goffard A, Behillil S, Enouf V, et al. Clinical features and viral diagnosis of two cases of infection with Middle East respiratory syndrome coronavirus: a report of nosocomial transmission. Lancet 2013;381:2265-2272. https://doi.org/10.1016/S0140-6736(13)60982-4
  18. Ehara N, Fukushima K, Kakeya H, Mukae H, Akamatsu S, Kageyama A, Saito A, Kohno S. A novel method for rapid detection of Streptococcus pneumoniae antigen in sputum and its application in adult respiratory tract infections. J Med Microbiol 2008;57:820-826. https://doi.org/10.1099/jmm.0.47793-0
  19. Sutantangjai M, Faksri K, Chaicumpar K, Chaimanee P, Lulitanond V, Namwat W. Evaluation of an immunochromatographic test kit for detecting Mycobacterium tuberculosis complex in sputum samples and on solid and in liquid cultures. Southeast Asian J Trop Med Public Health 2014;45:357-364.
  20. Matsuyama T, Morita T, Horikiri Y, Yamahara H, Yoshino H. Enhancement of nasal absorption of large molecular weight compounds by combination of mucolytic agent and nonionic surfactant. J Control Release 2006;110:347-352. https://doi.org/10.1016/j.jconrel.2005.09.047
  21. Saraswathy Veena V, Sara George P, Jayasree K, Sujathan K. Comparative analysis of cell morphology in sputum samples homogenized with dithiothreitol, N-acetyl-L cysteine, Cytorich® red preservative and in cellblock preparations to enhance the sensitivity of sputum cytology for the diagnosis of lung cancer. Diagn Cytopathol 2015;43:551-558. https://doi.org/10.1002/dc.23266
  22. Fischer AJ, Pino-Argumedo MI, Hilkin BM, Shanrock CR, Gansemer ND, Chaly AL, Zarei K, Allen PD, Ostedgaard LS, Hoffman EA, et al. Mucus strands from submucosal glands initiate mucociliary transport of large particles. JCI Insight 2019;4:e124863.
  23. Roberts GP. The role of disulfide bonds in maintaining the gel structure of bronchial mucus. Arch Biochem Biophys 1976;173:528-537. https://doi.org/10.1016/0003-9861(76)90289-7
  24. Carlson TL, Lock JY, Carrier RL. Engineering the mucus barrier. Annu Rev Biomed Eng 2018;20:197-220. https://doi.org/10.1146/annurev-bioeng-062117-121156
  25. Raj VS, Osterhaus AD, Fouchier RA, Haagmans BL. MERS: emergence of a novel human coronavirus. Curr Opin Virol 2014;5:58-62. https://doi.org/10.1016/j.coviro.2014.01.010
  26. Widagdo W, Begeman L, Schipper D, Run PR, Cunningham AA, Kley N, Reusken CB, Haagmans BL, van den Brand JM. Tissue distribution of the MERS-coronavirus receptor in bats. Sci Rep 2017;7:1193.
  27. Posthuma-Trumpie GA, Korf J, van Amerongen A. Lateral flow (immuno)assay: its strengths, weaknesses, opportunities and threats. A literature survey. Anal Bioanal Chem 2009;393:569-582. https://doi.org/10.1007/s00216-008-2287-2
  28. Li H, Han D, Hegener MA, Pauletti GM, Steckl AJ. Flow reproducibility of whole blood and other bodily fluids in simplified no reaction lateral flow assay devices. Biomicrofluidics 2017;11:024116.
  29. Miocevic O, Cole CR, Laughlin MJ, Buck RL, Slowey PD, Shirtcliff EA. Quantitative lateral flow assays for salivary biomarker assessment: a review. Front Public Health 2017;5:133.